HEPCIDIN, HEPCIDIN ANTAGONISTS AND METHODS OF USE

• US 20140286953A1
• Filed: 12/11/2013
• Published: 09/25/2014
• Est. Priority Date: 02/02/2007
• Status: Abandoned Application

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Sasu, Barbra, Haniu, Mitsuru, Boone, Thomas Charles, Bi, Xiao-juan, Lee, Grace Ki Jeong, Arvedson, Tara, Winters, Aaron George, Cooke, Keegan, Sheng, Jackie Z.
• Application Number:
  US14/103,351
• Publication Number:
  US 20140286953A1
• Time in Patent Office: Days
• US Class Current: 424/139.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 38/1816 Erythropoietin [EPO]
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 3/12 for electrolyte homeostasis
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/575 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 16/26 : against hormones ; against hormone releasing or inhibiting factors
• CPC Class Codes: C07K 2317/21 : from primates, e.g. man
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 15/1136 : against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/111 : spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/14 : interfering N.A.
• CPC Class Codes: C12N 2310/53 : partially self-complementary or closed
• CPC Class Codes: G01N 33/74 : involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

The invention relates to purified, correctly folded hepcidin, antibodies that bind hepcidin, and methods of making and using such materials. Also provide are methods of treated hepcidin-related disorders.

Claims

1. An isolated monoclonal antibody which specifically binds to human hepcidin, wherein said human hepcidin consists of the amino acid sequence set forth in SEQ ID NO:

9 and has a conformation comprising four disulfide-bond loops formed between residues 7 and 23, 10 and 13, 11 and 19, and 14 to 22 as located in SEQ ID NO;

9.

133. A composition comprising the antibody of claim 1 and pharmaceutically acceptable carrier, diluent or excipient.

134. A method for detecting the presence of human hepcidin in a biological sample comprising:

incubating the sample with the antibody of claim 1 under conditions that allow binding of the monoclonal antibody to mature human hepcidin of SEQ ID NO;

9; and

detecting the bound monoclonal antibody or bound hepcidin.

136. The method of claim 134, further comprising incubating the sample with a second monoclonal antibody that binds mature hepcidin of SEQ ID NO:

9.

172. An immunoassay kit for detecting human hepcidin in a biological sample, the kit comprising the antibody of claim 1 and, optionally purified hepcidin as a standard.

173. An immunoassay kit for detecting human hepcidin in a biological sample, comprising:

(a) a capture agent comprising an antibody that specifically binds to human hepcidin consisting of the amino acid sequence set forth in SEQ ID NO;

9; and

(b) a detection reagent comprising the antibody of claim 1 that binds to said human hepcidin at an epitope distinct from that recognized by the antibody in (a).

175. An immunoassay kit for detecting human hepcidin in a biological sample, comprising:

(a) a detection reagent comprising an antibody that specifically binds to human hepcidin consisting of the amino acid sequence set forth in SEQ ID NO;

9; and

(b) a capture reagent comprising the antibody of claim 1 that binds to said human hepcidin at an epitope distinct from that recognized by the antibody in (a).

179. A method of diagnosing a hepcidin-related disorder, comprising:

(a) contacting a biological sample from a human suspected of having said disorder with the antibody of claim 1 under conditions that allow binding of the antibody to human hepcidin; and

(b) detecting and/or quantitating the hepcidin bound to the antibody,

wherein the amount of hepcidin in said sample, as quantitated in (b), above a threshold level indicates the presence of hepcidin-related disorder and below the threshold level indicates the absence of hepcidin-related disorder.

181. The method according to claim 179, wherein said disorder is selected from the group consisting of:

anemia, sepsis, anemia of inflammation, anemia of cancer, chemotherapy induced anemia, chronic inflammatory anemia, congestive heart failure, end stage renal disorder, chronic kidney disease (stage I, II, III, IV or V), iron deficiency anemia, a disorder of iron homeostasis, ferroportin disease, hemochromatosis, diabetes, inflammation, rheumatoid arthritis, arteriosclerosis, tumors, vasculitis, systemic lupus erythematosus, hemoglobinopathies, and red cell disorders.

185. A method of monitoring a treatment in which a hepcidin antagonist is administered, comprising:

(a) contacting a biological sample, from a human that has been administered a hepcidin antagonist, with the antibody of claim 1 under conditions that allow binding of the antibody to human hepcidin; and

(b) detecting and/or quantitating the hepcidin bound to the antibody,

wherein the amount of hepcidin, as quantitated in (b), above a threshold level indicates that the dose of hepcidin antagonist is not therapeutically effective and below said threshold level indicates that the dose of hepcidin antagonist is therapeutically effective.

192. A vial or prefilled syringe comprising the composition of claim 1.

205. A kit for treating a disorder associated with elevated hepcidin levels or a disorder of iron homeostasis, comprising the antibody of claim 1 and an erythropoiesis stimulator.

210. A kit for treating a disorder associated with elevated hepcidin levels or a disorder of iron homeostasis, comprising the antibody of claim 1 and a label attached to or packaged with the container, the label describing use of the antibody with an erythropoiesis stimulator.

2-132. -132. (canceled)

135. (canceled)

137-171. -171. (canceled)

174. (canceled)

176-178. -178. (canceled)

180. (canceled)

182-184. -184. (canceled)

186-191. -191. (canceled)

193. A method of treating a mammal in need thereof comprising administering to said mammal (a) an antibody that binds human hepcidin consisting of SEQ ID NO:

9 with a K_D of about 10<sup>−

7</sup> M or less, and (b) an erythropoiesis stimulator selected from the group consisting of erythropoietin, erythropoietin variants and antibodies that bind erythropoietin.

194-196. -196. (canceled)

197. A method of treating a mammal that is hypo-responsive to therapy with an erythropoiesis stimulator, comprising administering an antibody that binds human hepcidin consisting of SEQ ID NO:

9 with a K_D of about 10<sup>−

7</sup>M or less.

198-204. -204. (canceled)

206-209. -209. (canceled)

211-236. -236. (canceled)

237. An isolated monoclonal antibody which specifically binds to human hepcidin, wherein said human hepcidin consists of the amino acid sequence set forth in SEQ ID NO:

9 and has a conformation comprising at least one disulfide-bond loop formed between residues 10 and 13 and 14 to 22 as located in SEQ ID NO;

9.